Comparative studies of four monoclonal antibodies to phenylalanine hydroxylase exhibiting different properties with respect to substrate-dependence, species-specificity and a range of effects on enzyme activity.
Four monoclonal antibodies to phenylalanine hydroxylase are described. Two are inhibitory (PH alpha 1-1 and PH alpha 2-1-1 antibodies), one is stimulatory (B5-1 antibody) and one has no effect on enzyme activity (PH alpha 3-0 antibody). Their properties are compared. Two antibodies (PH alpha 1-1 and B5-1 antibodies) bind primate and rodent phenylalanine hydroxylase, whereas the other two (PH alpha 2-1-1 and PH alpha 3-0 antibodies) bind only the primate enzyme. The binding of PH alpha 1-1 antibody to phenylalanine hydroxylase is dependent on substrate phenylalanine, whereas the binding of the others is not influenced by phenylalanine. Affinity adsorbents prepared from the four antibodies purified phenylalanine hydroxylase substantially (greater than 80% purity) in one step, except for a PH alpha 3-0 antibody--Sepharose column, which behaved anomalously. Two previous publications described the isolation and preliminary characterization of B5 and PH alpha 1-1 antibodies. PH alpha 2-1-1 and PH alpha 3-0 antibodies are reported for the first time.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000882:Haplorhini; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D051379:Mice; D010649:Phenylalanine / Q000378:metabolism; D010651:Phenylalanine Hydroxylase / Q000276:immunology*; D051381:Rats; D013045:Species Specificity; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D010649:Phenylalanine; D010651:Phenylalanine Hydroxylase